Part:BBa_K1763421
PCquad Mutant 10
One subunit of a 12-subunit protein cage with a thrombin cleavage sequence inserted between residues 243 and 248 (ALPSAE to ALPGLVPRGSGSAE).
This biobrick was designed to for controlled disassembly of a protein
cage in response to thrombin protease. What is a protein cage? A protein cage is a
type of supramolecular assembly, where multiple protein subunits interact non-
covalently in order to form a higher-order structure. Classic examples of protein
assemblies include viral capsids, ferritins, and clathrin coats. The cage that we are
working with is a previously-designed synthetic protein cage, created by
one of our advisors, Todd Yeates. To make a subunit of the cage, two distinct, naturally
oligomerizing proteins were selected—a trimer, bromoperoxidase, and a dimer, M1
matrix protein. Each were fused with a linker to form a 109.5 degree angle. This angle
is crucial in that it allows 12 subunits to come together, in order to create a cage that
has tetrahedral symmetry.
The goal of our project is to modify this protein cage such that disassembly can
be induced by thrombin protease. Proteases are enzymes that cleave at specific amino
acid sequences; therefore, if our cage contained the sequence shown here, it can be
disassembled by thrombin-induced cleavage. This would have potential applications
such as controlled release of loaded drugs/imaging molecules from inside the cage. So
why did we pick thrombin to work with? Thrombin was specifically chosen due to it
being relatively well-studied, and because of its role in the coagulation cascade, which
contributes to heart disease and stroke the first and fifth leading causes of death in the
United States, respectively.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 408
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 105
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 768
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